Team:Groningen/protocols/biofilm media

From 2013.igem.org

(Difference between revisions)
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<h5>Media for macrocolony formation</h5>
<h5>Media for macrocolony formation</h5>
-
<H5>2x SG medium (100 mL)</H5>
+
<H5>2x SG medium (100 ml)</H5>
<table id="normal" width="60%">
<table id="normal" width="60%">
   <tr>
   <tr>
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<tr>
<tr>
-
     <td>Ca(NO<sub>3</sub>)<sub>2</sub>.4H<sub>2</sub>O  1 mM</td>
+
     <td>Ca(NO<sub>3</sub>)<sub>2</sub>.4H<sub>2</sub>O  (1 mM)</td>
-
     <td>0.1 mL</td>
+
     <td>0.1 ml</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>MnCl<sub>2</sub>.4H<sub>2</sub>O 0.1 mM</td>
+
     <td>MnCl<sub>2</sub>.4H<sub>2</sub>O (0.1 mM)</td>
-
     <td>0.1 mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>FeSO<sub>4</sub>  1 &micro;M</td>
+
     <td>FeSO<sub>4</sub>  (1 &micro;M)</td>
-
     <td>0.1 mL</td>
+
     <td>0.1 ml</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
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<tr>
<tr>
     <td>0.1% glucose </td>
     <td>0.1% glucose </td>
-
     <td>0.5 mL</td>
+
     <td>0.5 ml</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
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-
<br><H5>LB/Mn/glucose medium (100 mL)</H5>
+
<br><H5>LB/Mn/glucose medium (100 ml)</H5>
<table id="normal" width="60%">
<table id="normal" width="60%">
   <tr>
   <tr>
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<tr>
<tr>
-
     <td>MnCl<sub>2</sub>.4H<sub>2</sub>O  0.1 mM</td>
+
     <td>MnCl<sub>2</sub>.4H<sub>2</sub>O  (0.1 mM)</td>
-
     <td>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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   <tr>
   <tr>
-
     <td>MOPS (100 mM)</td>
+
     <td>MOPS (100 mM)</td>
     <td>2.09 g</td>
     <td>2.09 g</td>
     <td></td>
     <td></td>
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   <tr>
   <tr>
-
     <td>MgCl<sub>2</sub>.6H<sub>2</sub>O  2 mM</td>
+
     <td>MgCl<sub>2</sub>.6H<sub>2</sub>O  (2 mM)</td>
     <td>0.04 g</td>
     <td>0.04 g</td>
     <td></td>
     <td></td>

Revision as of 00:08, 5 October 2013
Make an overnight culture of the strains of interest in LB broth.
1 ml of the O/N culture is required for every 10 ml of Sgg medium (2x SG supplemented with 1% [wt/vol] glycerol).
This will be incubated at 27°C for 5 days.
Media for macrocolony formation
2x SG medium (100 ml)
compound amount treatment
Nutrient broth (Difco) 1.6 g
KCl 0.2 g
MgSO4.7H2O 0.05 g
agar 1.5 g
add everything together and autoclave
Ca(NO3)2.4H2O (1 mM) 0.1 ml
MnCl2.4H2O (0.1 mM) 0.1 ml
FeSO4 (1 µM) 0.1 ml
0.1% glucose 0.5 ml

LB/Mn/glucose medium (100 ml)
compound amount treatment
LB/ 1.5% agar 100 ml autoclave
MnCl2.4H2O (0.1 mM) 0.1 ml
0.1% glucose 0.5 mL

MSgg medium (100 mL)
compound amount treatment
KH2PO4 0.026 g
K2HPO4 0.061 g
MOPS (100 mM) 2.09 g
MgCl2.6H2O (2 mM) 0.04 g
agar 1.5 g
add everything together and autoclave
CaCl2 (700 mM) 0.1 mL
MnCl2 (50 µM) 50 µL
FeCl3 (50 µM) 0.1 mL
ZnCl2 (1 µM) 0.1 mL
thiamine (2 µM) 0.1 mL
0.5% glycerol 0.57 mL
0.5% glutamate 10 mL
tryptophan 1 mL